Bacteriology of spoilage of fish muscle. IV. Role of protein.
Clarified muscle press juice from English sole (Parophrys vetulus) was fractionated by gel filtration into a protein and a protein-free fraction. Upon inoculation with spoilage bacteria, the protein fraction failed to show any signs of spoilage, but the protein-free fraction spoiled according to the usual organoleptic and chemical criteria. Despite its spoilage-resistant qualities, the protein fraction accelerated spoilage of the protein-free fraction when the two were combined. Protein breadkown due to bacterial action was greatest in the unfractionated juice and was least in the protein fraction. No significant proteolysis occurred until after spoilage became evident.


VARIATION IN DEVELOPMENTAL CONSTRAINTS IN SIGMODON.
Developmental constraints can be interpreted as factors of developmental origin responsible for covariation among measured variables. Several hypotheses have been proposed to link the possession of such constraints to subsequent evolution. Using confirmatory factor analysis, we compare developmental factors across selected taxa of cotton rats, genus Sigmodon. Three factors explain well the covariation among orofacial measurements: (1) responses to body size variation, (1) coordinated growth of traits of the occluding-tooth complex, and (3) responses to musculoskeletal interactions. Sigmodon taxa share these factors, but differ in the variance-covariance matrix of factors, and the unique variances of individual traits. Patterns of covariation among measurements of the neurocranial complex reflect responses to body size variation, and perhaps also responses to fetal brain growth. While there are no significant differences across taxa in factorpattern, variance-covariance matrix of factors, or unique variance of measured neurocranial variables, the neurocranium is only weakly constrained. We doubt that even the relatively stronger developmental constraints on the orofacial complex would prevent evolutionary divergence because differences in the variances and covariances of factors, and in levels of unique variance of individual traits can provide different opportunities for selection to act in different Sigmodon taxa.


Methoxy poly (ethylene glycol)-block-poly (glutamic acid)-graft-6-(2-nitroimidazole) hexyl amine nanoparticles for potential hypoxia-responsive delivery of doxorubicin.
Tumor microenvironment-responsive nano drug delivery vehicles are gaining mounting attention in the field of biomedical sciences. The hypoxic response of the tumorous cells due to very low partial pressure of oxygen (some time less than 2.5 mm of Hg) in the tumor tissues makes hypoxia-responsive drug delivery system as the more appealing in cancer chemotherapy. Based on these considerations, we synthesized hypoxia-responsive polymeric materials methoxy poly (ethylene glycol)-block-poly (glutamic acid)-graft-6-(2-nitroimidazole) hexyl amine (mPEG-b-PLG-g-NID) by conjugation of the hydrophobic nitro imidazole derivative (NID)[6-(2-nitroimidazole) hexyl amine] with the pendant carboxylic group of poly (ethylene glycol)-block-poly (L-glutamic acid)(mPEG-b-PLG). The structure and degree of substitution were confirmed by proton NMR, FTIR, and UV-Vis spectroscopy. The degree of substitution was found to enhance with the increase in NID to polymer ratio. The hypoxia response of the material was evaluated by UV-Vis spectroscopy and zeta potential measurements. Doxorubicin was hydrophobically encapsulated in the micellar core of the hypoxia-responsive nanoparticles. The drug-loaded micelles showed faster release in hypoxic condition as compared to normoxic conditions. Moreover, the developed polymeric system was found non-toxic to MCF-7 cell line, thus suggesting its biocompatibility and suitability as drug delivery device.


Mediastinal Hodgkin's disease--a retrospective analysis.
Thirty-seven patients with mediastinal Hodgkin's disease treated from 1972 to 1982 at the Division of Oncology of Basel University Hospital were analyzed retrospectively for their characteristics and the influence of various prognostic factors on survival. Deceased patients were older (35 vs. 25 years) and more often had systemic symptoms (8/22 vs. 3/15). 44% of the patients presented with mediastinal bulk disease, 66% of whom are in remission. No statistically significant influence of sex and stage was evident on the outcome. Patients treated with multimodality therapy had a significantly better survival than patients treated with radio- or chemotherapy only. No significant difference was found in survival in comparison to the population without mediastinal involvement.


Exploring the relationships between free-time management and boredom in leisure.
The purpose of the study was to examine the relations of five dimensions of free-time management (including goal setting and evaluating, technique, values, immediate response, and scheduling) with leisure boredom, and whether these factors could predict leisure boredom. A total of 500 undergraduates from a university in southern Taiwan were surveyed with 403 usable questionnaires was returned. Pearson correlation analysis revealed that five dimensions of free-time management had significant negative relationships with leisure boredom. Furthermore, the results of stepwise regression analysis revealed that four dimensions of free-time management were significant contributors to leisure boredom. Finally, we suggested students can avoid boredom by properly planning and organizing leisure time and applying techniques for managing leisure time.


A 12-bit high-speed column-parallel two-step single-slope analog-to-digital converter (ADC) for CMOS image sensors.
A 12-bit high-speed column-parallel two-step single-slope (SS) analog-to-digital converter (ADC) for CMOS image sensors is proposed. The proposed ADC employs a single ramp voltage and multiple reference voltages, and the conversion is divided into coarse phase and fine phase to improve the conversion rate. An error calibration scheme is proposed to correct errors caused by offsets among the reference voltages. The digital-to-analog converter (DAC) used for the ramp generator is based on the split-capacitor array with an attenuation capacitor. Analysis of the DAC's linearity performance versus capacitor mismatch and parasitic capacitance is presented. A prototype 1024 × 32 Time Delay Integration (TDI) CMOS image sensor with the proposed ADC architecture has been fabricated in a standard 0.18 μm CMOS process. The proposed ADC has average power consumption of 128 μW and a conventional rate 6 times higher than the conventional SS ADC. A high-quality image, captured at the line rate of 15.5 k lines/s, shows that the proposed ADC is suitable for high-speed CMOS image sensors.


Robotic radical hysterectomy is superior to laparoscopic radical hysterectomy and open radical hysterectomy in the treatment of cervical cancer.
Cervical cancer (CC) continues to be a global burden for women, with higher incidence and mortality rates reported annually. Many countries have witnessed a dramatic reduction in the prevalence of CC due to widely accessed robotic radical hysterectomy (RRH). This network meta-analysis aims to compare intraoperative and postoperative outcomes in way of RRH, laparoscopic radical hysterectomy (LTH) and open radical hysterectomy (ORH) in the treatment of early-stage CC. A comprehensive search of PubMed, Cochrane Library and EMBASE databases was performed from inception to June 2016. Clinical controlled trials (CCTs) of above three hysterectomies in the treatment of early-stage CC were included in this study. Direct and indirect evidence were incorporated for calculating values of weighted mean difference (WMD) or odds ratio (OR), and drawing the surface under the cumulative ranking curve (SUCRA). Seventeen 17 CCTs were ultimately enrolled in this network meta-analysis. The network meta-analysis showed that patients treated by RRH and LRH had lower estimated blood loss compared to patients treated by ORH (WMD = -399.52, 95% CI = -600.64~-204.78; WMD = -277.86, 95%CI = -430.84 ~ -126.07, respectively). Patients treated by RRH and LRH had less hospital stay (days) than those by ORH (WMD = -3.49, 95% CI = -5.79~-1.24; WMD = -3.26, 95% CI = -5.04~-1.44, respectively). Compared with ORH, patients treated with RRH had lower postoperative complications (OR = 0.21, 95%CI = 0.08~0.65). Furthermore, the SUCRA value of three radical hysterectomies showed that patients receiving RRH illustrated better conditions on intraoperative blood loss, operation time, the number of resected lymph nodes, length of hospital stay and intraoperative and postoperative complications, while patients receiving ORH demonstrated relatively poorer conditions. The results of this meta-analysis confirmed that early-stage CC patients treated by RRH were superior to patients treated by LRH and ORH in intraoperative blood loss, length of hospital stay and intraoperative and postoperative complications, and RRH might be regarded as a safe and effective therapeutic procedure for the management of CC.


Cardiac rhabdomyoma in infancy. A case report.
We describe a case of rhabdomyoma of the heart in a newborn infant and present an overview of current knowledge about the natural history of these lesions, with implications for management. This is the only case of its kind seen at this hospital.


Interrelations between the small isolated groups with homogeneous and heterogeneous composition.
Increase in the heterogeneity of a space crew's composition (cultural and gender differences) is a risk factor that can negatively influence the formation of a cohesive crew, which depends on a common way of perceiving one's social environment, goals and values. Method. Three groups working in isolation from 110 to 240 days, all with different cultural and gender composition were analysed using a modification of Kelly's repertory grid technique during a space simulation at the Institute for Biomedical Problems, Moscow. Subjects assessed themselves and each other monthly. Results. National Russian and international groups failed to create a single cohesive crew. Members of both groups considered people from their group as "us" and their neighbours as "them". Their relations became more negative during the mission, with attitudes changing from neutral to poor. Subjects began to perceive each other as "different and distant". Cultural and language differences prevented national (Russian) and international groups from forming a common understanding of behavioural rules and establishing close emotional contacts. Conclusions. Differences in perception of the interpersonal environment, group goals and values in national and international groups hindered the formation of a joint cohesive crew.


Effect of restoration of euthyroidism on peripheral blood cells and erythropoietin in women with subclinical hypothyroidism.
In overt hypothyroidism (OH) anemia is common, while less frequently basophilia has been described. In subclinical hypothyroidism (SCH), however, data on the distribution of peripheral blood cells are lacking. Therefore, we evaluated the effects of L-T4 replacement therapy on peripheral blood elements in female patients with SCH before and after restoration of euthyroidism in a randomized, double-blind and placebo-controlled study. Sixty-six women with SCH (TSH 12.9 +/- 8.2 mU/L) were randomly assigned to receive L-thyroxine or placebo for 48 weeks. 63 of the 66 women completed the study. Peripheral blood cells were measured at baseline and 48 weeks after L-thyroxine or placebo treatment, respectively. The percentage of lymphocytes decreased (p<0.05), whereas percent of monocytes (p<0.05) and eosinophiles (p<0.05) increased significantly upon restoration of euthyroidism after 48 weeks. Hemoglobin and hematocrit remained unchanged throughout the study period. However, erythropoietin levels increased significantly (p<0.01) during L-T4 treatment. In the placebo group all parameters remained unchanged throughout the study. Overall, we observed subtle alterations of the leuco-lympho-monocytic distribution of the peripheral blood cells upon restoration of euthyroidism in patients with SCH. Hemoglobin and Hematocrit remained unchanged; however, the increasing level of erythropoietin during L-T4 treatment suggests an already stimulated, yet compensated erythropoietic system in mild thyroid failure.


Relationship between uncertainty in the oil and stock markets before and after the shale gas revolution: Evidence from the OVX, VIX, and VKOSPI volatility indices.
We investigate the relationship between crude oil prices and stock markets. Unlike prior studies, we use implied volatility indices and evaluate the change in the relationship between the volatility indices through a sub-period analysis. Specifically, we examine the causal relationships among the crude oil, S&P 500 index, and KOSPI 200 index volatilities by using the autoregressive distributed lag (ARDL) bounds and the Toda-Yamamoto Granger causality tests. In addition, a BEKK-GARCH model is employed to enhance the robustness of the causality test results. These experiments indicate that the OVX and VIX show bi-directional causality in the period that includes the shale gas revolution and no causality in the period that does not. Further, the OVX Granger causes the VKOSPI in the former period, but there is no causality between them in the latter period. Finally, we find strong unidirectional causality from the VIX to the VKOSPI in both sub-periods. These results have important implications for the analysis of portfolio risk management and for assisting energy policymakers and traders in making effective decisions and investments, respectively.


[Fertility with uterine anomalies].
45 patients with uterine anomalies were studied regarding their fertility. 39 of these 45 patients were pregnant prior or following the diagnosis. The 80 conceptions show a fertility rate of 86.6%. 46 pregnancies led to the delivery of a viable infant. 34 pregnancies resulted in miscarriage of premature delivery. Miscarriages were much more frequent in women with bicornuate or unicornuate uteri (60%). When other possible reasons for abortions such as toxoplasmosis, insufficiency of the cervix or deficient luteal phase are corrected the number of pregnancies rose to 82. In 53 of these cases a viable infant was delivered. 30% of the women had to be delivered by Caesarean section. When the desire for child-bearing exists uterine anomalies need to be first treated conservatively. Surgical correction of the anomaly is only indicated in a limited number of these cases.


Predation of endangered Arctic foxes by Golden eagles: What do we know?
Dedicated conservation efforts spanning the past two decades have saved the Fennoscandian Arctic fox (Vulpes lagopus) population from local extinction, and extensive resources continue to be invested in the species' conservation and management. Although increasing, populations remain isolated, small and are not yet viable in the longer term. An understanding of causes of mortality are consequently important to optimize ongoing conservation actions. Golden eagles (Aquila chrysaetos) are a predator of Arctic foxes, yet little information on this interaction is available in the literature. We document and detail six confirmed cases of Golden eagle depredation of Arctic foxes at the Norwegian captive breeding facility (2019-2022), where foxes are housed in large open-air enclosures in the species' natural habitat. Here, timely detection of missing/dead foxes was challenging, and new insights have been gained following recently improved enclosure monitoring. Golden eagle predation peaked during the winter months, with no cases reported from June to November. This finding contrasts with that which is reported from the field, both for Arctic and other fox species, where eagle depredation peaked at dens with young (summer). While the seasonality of depredation may be ecosystem specific, documented cases from the field may be biased by higher survey efforts associated with the monitoring of reproductive success during the summer. Both white and blue color morphs were housed at the breeding station, yet only white foxes were preyed upon, and mortality was male biased. Mitigation measures and their effectiveness implemented at the facility are presented. Findings are discussed in the broader Arctic fox population ecology and conservation context.


[Succinylcholine use by anesthesiologists in Croatia--is it really abandoned?].
The aim was to establish the prevalence of succinylcholine use among Croatian anesthesiologists in adult elective and emergency surgery, as well as in pediatric surgery, regarding gender, position, working place, and working experience of physicians. The anesthesiologists were expected to express their personal opinions regarding the drug, as well as experienced side effects in their own clinical practice. A total of 125 anesthesiologists (out of 590 in Croatia) from both university and county hospitals in Croatia anonymously filled out the questionnaire regarding the use of succinylcholine (Appendix 1). The questionnaire was structured to assess the use of succinylcholine in adult elective and emergency surgery, and in pediatric anesthesia, to obtain the reasons for the preference or rejection of succinylcholine, and information about observed side effects. The differences in use regarding gender, position, working place, and working experience were tested using chi-squared test and Fisher's exact test. p < 0.05 was considered significant. Vast majority (approximately 70%) of anesthesiologists in Croatia still use succinylcholine. The percentages of anesthesiologists that never use succinylcholine in adult elective, adult emergency and pediatric surgery were 20%, 6%, and 31%, respectively. There were no significant differences in the use of succinylcholine regarding position, working place, and working experience, but male anesthesiologists used it less frequently in pediatric anesthesia compared with their female colleagues (chi2 = 5.08; p = 0.02). Forty-two per cent never experienced a complication from the drug use. The most frequently reported side effects were bradycardias (67%) and myalgias (54%), followed by prolonged blockade (33%), and allergy (33%). Asystole was reported by 10% of the respondents. In conclusion, succinylcholine is still widely used by anesthesiologists in Croatia. The majority of surveyed physicians were aware of its possible dangerous adverse effects, but still use it in certain situations. Therefore, indications and contraindications for its use deserve expert consensus guidelines based on the available scientific evidence.


Rheumatic fever: update on the Jones criteria according to the American Heart Association review - 2015.
Rheumatic fever is still currently a prevalent disease, especially in developing countries. Triggered by a Group A β-hemolytic Streptococcus infection, the disease may affect genetically predisposed patients. Rheumatic carditis is the most important of its clinical manifestations, which can generate incapacitating sequelae of great impact for the individual and for society. Currently, its diagnosis is made based on the Jones criteria, established in 1992 by the American Heart Association. In 2015, the AHA carried out a significant review of these criteria, with new diagnostic parameters and recommendations. In the present study, the authors perform a critical analysis of this new review, emphasizing the most relevant points for clinical practice.


Spontaneous recurrence of a summer-like diel rhythm in the body temperature of the Syrian hamster after hibernation.
Mammalian hibernation is a survival strategy characterized by metabolic suppression and drastically lowering body temperature (Tb), used during harsh seasons with food shortages and cold. The Syrian hamster commences hibernation in response to a short photoperiod and cold but spontaneously concludes hibernation after several months without environmental cues. Little is known about the changes in diel rhythms during hibernation. Using long-term and high-resolution Tb data, we analysed the diel Tb rhythm time-course changes in Syrian hamsters raised under summer-like conditions (long photoperiod (LP) and warm; LP-warm) and transferred to winter-like conditions (short photoperiod (SP) and cold; SP-cold). The diel Tb rhythm was undetectable during the hibernation period (HIBP), reappearing after the HIBP. The phase of this returning rhythm reverted to the LP entrainment phase characteristics despite the ambient SP and then re-entrained to the ambient SP as if the hamsters were transferred from the LP-warm to SP-cold conditions. The diel Tb rhythm reverted from the SP- to LP-type in a hibernation-dependent manner. Under constant dark and cold conditions, the circadian Tb rhythm recovered without photic stimuli following the HIBP. These findings suggest that hibernation involves a program that anticipates the ambient photoperiod when animals emerge from hibernation.


Quantitative assessment of Na+,K+-ATPase localization by direct and indirect p-nitrophenyl phosphatase methods.
Na+,K+-ATPase histochemistry, using direct (Pb) and indirect (Mg/Sr-Co) p-nitrophenyl phosphatase methods, was assessed by scanning integrative microdensitometry of three classes of tubules in mouse and guinea pig kidneys. The methods yielded similar and appropriate patterns of activity distribution and inhibitor response. The indirect method gave preferable results, in that non-enzymic background was lower and rate of reaction product accumulation was considerably higher.


Youth ice hockey COVID-19 protocols and prevention of sport-related transmission.
This observational study evaluated the impact of return-to-play protocols to prevent transmission of SARS-CoV-2 in a youth ice hockey programme in Virginia Beach, Virginia. Following an outbreak of SARS-CoV-2 in November 2020, a COVID-19 Response Team evaluated the epidemiological data to identify transmission dynamics and develop enhanced protocols to prevent transmission. During the subsequent 18-week study period, incident cases were investigated to identify the likely transmission source; testing, quarantine and isolation recommendations were provided to families in accordance with Centers for Disease Control and Prevention guidelines. Simple but stringent protocols were implemented among 148 youth ice hockey players ages 6-18. Players were required to arrive at the rink in full gear; locker rooms were closed, building entry was limited to one parent per player, and masks were required at all times except for players on the ice. Following implementation of the enhanced protocols, more than 500 practices and games were completed with at least 15 858 athlete-hours of exposure and no within-programme COVID-19 transmission was detected despite high community incidence and sporadic household exposures. This study suggests indoor youth sports can operate safely with appropriate protocols in place, even within communities of high COVID-19 transmission, even when athletes are not yet vaccinated or wearing masks during play. Transmission appears to be more likely in congested indoor areas involving adults than on the ice during play. Protocols should be developed in collaboration with programme participants. Strong collaboration in the interest of youth sports can motivate adoption of protocols which prevent within-team transmission.


UHPLC-MS phytochemical profiling, biological propensities and in-silico studies of Alhagi maurorum roots: a medicinal herb with multifunctional properties.
The biological, chemical, and in silico properties of methanol and dichloromethane (DCM) extracts of Alhagi maurorum roots with respect to the antioxidant, enzyme inhibition, and phytochemical composition were evaluated. Total bioactive contents were determined spectrophotometrically, and the individual secondary metabolites composition was assessed via ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) analysis. Antioxidant capacities were evaluated using a panoply of assays (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical scavenging, ferric reducing antioxidant power (FRAP), cupric reducing antioxidant power (CUPRAC), phosphomolybdenum total antioxidant capacity (TAC), and metal chelating activity (MCA)). The enzyme inhibition potential was studied against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase, tyrosinase, urease and lipoxygenase (LOX) enzymes. The methanol extract was found to contain higher total phenolic (105.91 mg GAE/g extract) and flavonoid (2.27 mg RE/g extract) contents which can be correlated to its more substantial antioxidant potential as well as AChE, BChE, tyrosinase and α-glucosidase inhibition. However, the DCM extract was the most effective against α-amylase (1.86 mmol ACAE/g extract) enzyme inhibition. The UHPLC-MS analysis of methanol extract identified the tentative presence of a total of 18 secondary metabolites, including flavonoids, saponins, phenolic and terpenoid derivatives. Three compounds named emmotin A, luteolin 5,3'-dimethyl ether, and preferrugone were further investigated for their in silico molecular docking studies against the tested enzymes. The selected compounds were found to have higher binding interaction with AChE followed by BChE, α-glucosidase, α-amylase, and tyrosinase. The results of the present study have demonstrated A. mauroram to be considered as a lead source of natural antioxidant and enzyme inhibitor compounds.


Non-Hodgkin's lymphoma with rheumatism-like presentation and multiple cystic and nodular lesions in the liver.
We report the case of a non-Hodgkin's lymphoma in an isolated and not previously diagnosed "adult polycystic liver disease". In this case, a rheumatism-like presentation was associated with the discovery of multiple nodular and cystic lesions in the liver. Of these, the former were related to lymphoma involvement, the latter to pre-existent liver disease. The singularity of the case, not previously reported, and some difficulties met in diagnosing the two pathologies, justify the report.


Kinetics of polymer looping with macromolecular crowding: effects of volume fraction and crowder size.
The looping of polymers such as DNA is a fundamental process in the molecular biology of living cells, whose interior is characterised by a high degree of molecular crowding. We here investigate in detail the looping dynamics of flexible polymer chains in the presence of different degrees of crowding. From the analysis of the looping-unlooping rates and the looping probabilities of the chain ends we show that the presence of small crowders typically slows down the chain dynamics but larger crowders may in fact facilitate the looping. We rationalise these non-trivial and often counterintuitive effects of the crowder size on the looping kinetics in terms of an effective solution viscosity and standard excluded volume. It is shown that for small crowders the effect of an increased viscosity dominates, while for big crowders we argue that confinement effects (caging) prevail. The tradeoff between both trends can thus result in the impediment or facilitation of polymer looping, depending on the crowder size. We also examine how the crowding volume fraction, chain length, and the attraction strength of the contact groups of the polymer chain affect the looping kinetics and hairpin formation dynamics. Our results are relevant for DNA looping in the absence and presence of protein mediation, DNA hairpin formation, RNA folding, and the folding of polypeptide chains under biologically relevant high-crowding conditions.


Cervical Alpha-Actinin-4 Is Upregulated in Women with Threatened Preterm Labor and Microbial Invasion of the Amniotic Cavity.
To characterize the proteome profile of women with threatened preterm labor (PTL) below 34;0 weeks with and without microbial invasion of the amniotic cavity (MIAC) using mass spectrometry in the amniotic fluid (AF) and Western blot analysis in the cervical mucus and the vaginal fluid. In the discovery phase, a case-control study including 8 women with MIAC and 7 without matched for gestational age at sampling was performed. Proteomic profile characterization was done using the LTQ VELOS Orbitrap mass spectrometer in the AF. In the validation phase, a selection of the proteins differentially expressed by mass spectrometry in the genital samples of a prospective cohort of 109 women was validated by Western blot analysis. In the discovery phase, the mass spectrometry analysis identified a total of 444 proteins. Sixteen were chosen for validation, being involved in defense (calgranulin A, B, C, C-reactive protein), cytoskeletal remodeling (alpha-actinin-4 [ACTN-4], plastin-2, α2-antiplasmin, vitronectin), metabolism (cystatin-β, glucose 6 phosphate isomerase, glutathione S-transferase, prostaglandin D2 synthase, corticosteroid-binding globulin), and vascular (α1-antichymotrypsin, hemopexin, endosialin) pathways. In the validation phase, cervical ACTN-4 was the only significantly upregulated protein in women with MIAC with an odds ratio of 6.8 (p = 0.002). Cervical ACTN-4 was significantly upregulated in the group of women with PTL with MIAC.


The tobacco cembranoid (1S,2E,4S,7E,11E)-2,7,11-cembratriene-4,6-diol as a novel angiogenesis inhibitory lead for the control of breast malignancies.
(1S,2E,4S,6R,7E,11E)-2,7,11-cembratriene-4,6-diol (1) and its 4-epi-analog (2) are diterpene precursors of the key flavor components in most Nicotiana (tobacco) species that purposely degraded during commercial tobacco fermentation. Angiogenesis, recruitment of new blood vessels, is important for tumor growth, survival and metastasis that can be targeted to control cancer. This study shows evidences and potential of the cembranoid 1 as a potent angiogenesis modulator through targeting VEGFR2. In silico study suggested favorable docking scores and binding affinity of 1 at the ATP binding pocket of VEGFR2. The binding mode of 1 was parallel to the standard FDA-approved antiangiogenic drug sunitinib (4). In vitro, cembranoid 1 significantly reduced the activated VEGFR2 levels in multiple breast cancer cell lines. Intraperitoneal 40mg/kg, 3X/week treatment of 1 significantly reduced the MDA-MB-231 cells breast tumor size in mice. Immunohistochemistry and Western blotting analysis of the treated mice tumors showed significant downregulation of the vasculogenesis marker CD31 and suppressed activated VEGFR2-paxillin-FAK pathway. Matrigel study in Swiss albino mice showed similar trend. The tobacco cembranoid 1 is a potential antiangiogenic lead useful for future use to control breast malignancies.


Anti-platelet drugs: do they affect megakaryocytes?
Anti-platelet drugs are used in clinical medicine to prevent thromboembolic complications of cardiovascular diseases. Among anti-platelet drugs, very little is known of their possible effects on megakaryocytes. ASA is the only compound for which it has clearly been demonstrated that its mechanism of action involves acetylation of the Ser 529 residue in cyclo-oxygenase in platelets and megakaryocytes. Because megakaryocytes possess membrane receptors for ADP, the thienopyridine metabolites of ticlopidine and clopidogrel may modify these receptors as in platelets and hence prevent ADP binding and further activation. Megakaryocytes also have GPIIb-IIIa receptors for the adhesive protein fibrinogen and may be accessible in vivo to GPIIb-IIIa antagonists such as the monoclonal antibody abciximab. Drugs such as heparin or the phosphodiesterase inhibitor anagrelide can either inhibit or stimulate megakaryocytopoiesis and platelet production, while cytokines such as thrombopoietin affect megakaryocytopoiesis, platelet production and platelet function by potentiating the activation of platelets by other agonists.


Prognosis of metastatic gastric and gastroesophageal junction cancer by HER2 status: a European and USA International collaborative analysis.
To determine whether human epidermal growth factor receptor 2 (HER2) status is an independent prognostic factor in metastatic gastric and gastroesophageal junction (GEJ) adenocarcinoma. Formalin-fixed paraffin-embedded tumor samples from 381 metastatic gastric/GEJ cancer patients enrolled at Krankenhaus Nordwest and Memorial Sloan-Kettering Cancer Centers on six first-line trials of chemotherapy without trastuzumab were examined for HER2 by immunohistochemistry (IHC) and in situ hybridization (ISH). IHC 3+ or ISH-positive tumors were considered HER2 positive. Seventy-eight of 381 patients (20%) had HER2-positive disease. In the multivariate logistic model, there were significantly higher rates of HER2 positivity in patients with liver metastasis (liver metastasis 31%; no liver metastasis 11%; P = 0.025) and intestinal histology (intestinal 33%; diffuse/mixed 8%; P = 0.001). No significant differences in HER2 positivity were found between resections and biopsies or primaries and metastases. Patients with HER2-positive gastric cancer had longer median overall survival compared with HER2-negative gastric cancer patients (13.9 versus 11.4 months, P = 0.047), but multivariate analysis indicated that HER2 status was not an independent prognostic factor (hazard ratio 0.79; 0.44-1.14; P = 0.194). Approximately 20% of Western patients with metastatic gastric cancer are HER2 positive. Unlike breast cancer, HER2 positivity is not independently prognostic of patient outcome in metastatic gastric or GEJ.


Evaluation of Patient Motivation and Satisfaction During Technology-Assisted Rehabilitation: An Experiential Review.
Objective: The aim of this experiential review is to explore the state of the art of the literature regarding the evaluation tools available for assessment of patient motivation and satisfaction during technology-assisted rehabilitation (robot rehabilitation, virtual reality rehabilitation, and serious games rehabilitation). Materials and Methods: A systematic search of the peer-reviewed literature published from January 1990 to August 2019 was conducted. The protocol for this review was registered in PROSPERO and carried out in accordance with the PRISMA recommendations. Results: The search of PubMed, PsycINFO, Scopus, and Web of Science databases identified a total of 333 records. After adjusting for duplicates and other inclusion criteria, 69 studies were selected for inclusion in the review. We found that authors used a wide range of dedicated questionnaires and, in about 50% of studies, a few validated tools to assess motivation and satisfaction during technology-assisted rehabilitation. The instruments most used were the Intrinsic Motivation Inventory (IMI), Quebec User Evaluation of Satisfaction with Assistive Technology (QUEST 2.0), and the Usefulness, Satisfaction, and Ease of use (USE) scale. Motivation and satisfaction were generally portrayed as multidimensional concepts; overall, 29 domains were assessed by 9 different tools. Conclusion: The tools used in the current literature to assess patient motivation and satisfaction during technology-assisted rehabilitation are quite variegated, but we would recommend use of the IMI and USE questionnaires based on their widespread diffusion. However, the choice of domains explored and number of items calls for harmonization. Ideally, this should be a joint task for the whole scientific community.


The fractionation of spoken language understanding by measuring electrical and magnetic brain signals.
This paper focuses on what electrical and magnetic recordings of human brain activity reveal about spoken language understanding. Based on the high temporal resolution of these recordings, a fine-grained temporal profile of different aspects of spoken language comprehension can be obtained. Crucial aspects of speech comprehension are lexical access, selection and semantic integration. Results show that for words spoken in context, there is no 'magic moment' when lexical selection ends and semantic integration begins. Irrespective of whether words have early or late recognition points, semantic integration processing is initiated before words can be identified on the basis of the acoustic information alone. Moreover, for one particular event-related brain potential (ERP) component (the N400), equivalent impact of sentence- and discourse-semantic contexts is observed. This indicates that in comprehension, a spoken word is immediately evaluated relative to the widest interpretive domain available. In addition, this happens very quickly. Findings are discussed that show that often an unfolding word can be mapped onto discourse-level representations well before the end of the word. Overall, the time course of the ERP effects is compatible with the view that the different information types (lexical, syntactic, phonological, pragmatic) are processed in parallel and influence the interpretation process incrementally, that is as soon as the relevant pieces of information are available. This is referred to as the immediacy principle.


Health inequalities in the Caribbean: increasing opportunities and resources.
Social inequalities in health are not a priority in the Caribbean region. However, a few studies suggested such disparities and indicators show important socioeconomic disparities between and within countries. There are indications that governments' investment in health and other social programs is insufficient in the region and that regional health institutions that guide national health policies and programs do not make the reduction of social inequalities a priority. Furthermore, the public health sector is generally weak and health services are mainly focusing on curative services. The author argues that there is a need to develop and to implement social policies that include equity and social justice as core values. In order to increase the focus on health inequalities in the region, there is also a need to strengthen the Public Health field that integrates Health Promotion strategies. It is also suggested that international, regional and national health sectors that include academic and research institutions, health-related journals and associations, and non-governmental organizations put health inequalities in the Caribbean on their agenda. Furthermore, there should be a fundamental switch from a biomedical perspective of health to a paradigm that considers health as the expression of political, social and economic circumstances.


Two Biexciton Types Coexisting in Coupled Quantum Dot Molecules.
Coupled colloidal quantum dot molecules (CQDMs) are an emerging class of nanomaterials, manifesting two coupled emission centers and thus introducing additional degrees of freedom for designing quantum-dot-based technologies. The properties of multiply excited states in these CQDMs are crucial to their performance as quantum light emitters, but they cannot be fully resolved by existing spectroscopic techniques. Here we study the characteristics of biexcitonic species, which represent a rich landscape of different configurations essentially categorized as either segregated or localized biexciton states. To this end, we introduce an extension of Heralded Spectroscopy to resolve the different biexciton species in the prototypical CdSe/CdS CQDM system. By comparing CQDMs with single quantum dots and with nonfused quantum dot pairs, we uncover the coexistence and interplay of two distinct biexciton species: A fast-decaying, strongly interacting biexciton species, analogous to biexcitons in single quantum dots, and a long-lived, weakly interacting species corresponding to two nearly independent excitons. The two biexciton types are consistent with numerical simulations, assigning the strongly interacting species to two excitons localized at one side of the quantum dot molecule and the weakly interacting species to excitons segregated to the two quantum dot molecule sides. This deeper understanding of multiply excited states in coupled quantum dot molecules can support the rational design of tunable single- or multiple-photon quantum emitters.


Cryotherapy Treatment After Arthroscopic Knee Debridement and ACL Reconstruction: A Review.
Pain, swelling, and inflammation of the knee joint and its surrounding soft-tissue structures are common during the postoperative period after arthroscopic knee debridement and anterior cruciate ligament reconstruction. These challenges can make patient recovery difficult immediately after surgery. Several options exist, however, to help patients overcome these challenges. Cryotherapy has been noted to decrease pain, swelling, and inflammation. However, while a number of studies exist characterizing the use of cryotherapy after knee surgery, no definitive cryotherapy devices and modalities have been identified. Therefore, the purpose of this review was to evaluate randomized controlled trials to assess the use of cryotherapy after: 1) arthroscopic debridement; and 2) anterior cruciate ligament reconstruction.


Spatially Controlled Occlusion of Polymer-Stabilized Gold Nanoparticles within ZnO.
In principle, incorporating nanoparticles into growing crystals offers an attractive and highly convenient route for the production of a wide range of novel nanocomposites. Herein we describe an efficient aqueous route that enables the spatially controlled occlusion of gold nanoparticles (AuNPs) within ZnO crystals at up to 20 % by mass. Depending on the precise synthesis protocol, these AuNPs can be (i) solely located within a central region, (ii) uniformly distributed throughout the ZnO host crystal or (iii) confined to a surface layer. Remarkably, such efficient occlusion is mediated by a non-ionic water-soluble polymer, poly(glycerol monomethacrylate)70 (G70 ), which is chemically grafted to the AuNPs; pendent cis-diol side groups on this steric stabilizer bind Zn2+ cations, which promotes nanoparticle interaction with the growing ZnO crystals. Finally, uniform occlusion of G70 -AuNPs within this inorganic host leads to faster UV-induced photodegradation of a model dye.


Preclinical models of pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDA) is one of the most difficult human malignancies to treat. The 5-year survival rate of PDA patients is 7% and PDA is predicted to become the second leading cancer-related cause of death in the USA. Despite intensive efforts, the translation of findings in preclinical studies has been ineffective, due partially to the lack of preclinical models that faithfully recapitulate features of human PDA. Here, we review current preclinical models for human PDA (eg human PDA cell lines, cell line-based xenografts and patient-derived tumour xenografts). In addition, we discuss potential applications of the recently developed pancreatic ductal organoids, three-dimensional culture systems and organoid-based xenografts as new preclinical models for PDA.


Characterization of PTEN mutations in brain cancer reveals that pten mono-ubiquitination promotes protein stability and nuclear localization.
PTEN is a PIP3 phosphatase that antagonizes oncogenic PI3-kinase signalling. Due to its critical role in suppressing the potent signalling pathway, it is one of the most mutated tumour suppressors, especially in brain tumours. It is generally thought that PTEN deficiencies predominantly result from either loss of expression or enzymatic activity. By analysing PTEN in malignant glioblastoma primary cells derived from 16 of our patients, we report mutations that block localization of PTEN at the plasma membrane and nucleus without affecting lipid phosphatase activity. Cellular and biochemical analyses as well as structural modelling revealed that two mutations disrupt intramolecular interaction of PTEN and open its conformation, enhancing polyubiquitination of PTEN and decreasing protein stability. Moreover, promoting mono-ubiquitination increases protein stability and nuclear localization of mutant PTEN. Thus, our findings provide a molecular mechanism for cancer-associated PTEN defects and may lead to a brain cancer treatment that targets PTEN mono-ubiquitination.


[Differential effects on bone and mesenchymal stem cells caused by intermittent and continuous PTH administration].
Objective: To investigate the distinct effects of intermittent and continuous administration of parathyroid hormone (PTH) on bone and mesenchymal stem cell (MSC). Methods: Six weeks old mice with C57/BL6J background and SOX9-creERT/Td-tomato/Osteocalcin-GFP genotype were divided into 6 groups: intermittent administration and withdraw group (subcutaneous injection with PTH 500 μg·kg-1·d-1), continuous administration and withdraw group (subcutaneous implantation of PTH pump, 80 μg·kg-1·d-1, with a rate of 0.25 μl/h), control administration and withdraw group, with 8 mice in each group. Serum calcium level and bone mineral density (BMD) were measured after two weeks' treatment and two weeks after drug withdraw. Histopathology and immunofluorescence analyses were performed to assess the effects of PTH on bone and mesenchymal stem cell. Results: Serum calcium level increased transiently in intermittent group[(1.36±0.03) mmol/L]and increased gradually in continuous group[up to (2.33±0.03) mmol/L], but reduced to normal level (1.12-1.27 mmol/L) 14 days after drug withdraw. BMD of both intermittent[(0.047±0.002) g/cm2]and continuous[(0.046±0.001) g/cm2]PTH administration groups increased compared with control group[(0.044±0.001) g/cm2], but there was no significant difference among three groups 2 weeks after drug withdraw. Femoral histopathology showed that bone mass, trabecular number and little fibrous tissue hyperplasia in intermittent PTH group increased. Osteoblasts number increased, but lining cells decreased. There was no significant difference in osteocyte and osteoclast numbers. After withdrawing of intermittent PTH, osteocyte and osteoblast number declined significantly, but there was an increased number of lining cells. Continuous PTH caused very high amount of fibrosis, and osteoclast number increased significantly, while osteoblast and osteocyte number increased slightly. After withdrawing of continuous PTH, fibrosis disappeared significantly, and lining cell number increased. Immunofluorescence staining in the epiphyseal-metaphyseal regions in fibula showed intermittent PTH increased undifferentiated Td-Tomato MSC, but declined significantly after withdrawing. Undifferentiated Td-Tomato MSC in continuous PTH increased slightly and decreased after drug withdraw. Conclusions: Intermittent PTH increased undifferentiated Td-Tomato MSC and osteoblast number, and might transform lining cell into osteocytes and osteoblasts, and thus lead to bone formation. Continuous PTH increased undifferentiated Td/Tomato MSC, osteoblast and osteocyte number slightly, but high amount of fibrosis and osteoclasts can be seen, leading to metabolic bone disease. However, lining cell ascended after drug withdraw, which may be beneficial to bone remodeling.


Immediate aftermath of a client's death: the experience of home health aides.
This study investigated Home Health Aides' (HHAs) experiences in the immediate aftermath of client death. Semi-structured in-person interviews with 80 HHAs explored how notification of death and reassignment to a new client were handled. Only 42.5% of HHAs were notified of the death; 40% had to notify the agency; 17.5% were not notified at all and had a negative experience. Reassignment preferences varied, but HHAs had a better experience when their preferences were taken into consideration. Study findings suggest that more mindful approaches to transitions following client death would be valued by HHAs and could improve their work experience.


How will B-cell-receptor-targeted therapies change future CLL therapy?
For many years there has been considerable disassociation between the understood biology of chronic lymphocytic leukemia (CLL) and the therapeutics used to treat this disease. With the introduction of the first targeted CD20 antibody rituximab and its addition to chemotherapy came the first observation that minimal residual disease-negative (MRD-negative) complete responses (CRs) could be obtained with dramatically improved progression-free survival and overall survival. This advance was soon to be surpassed by the introduction of therapeutics that target B-cell receptor (BCR) signaling. New data show that BCR-inhibiting agents are very active for the treatment of relapsed CLL, despite the lack of MRD-negative CR, with durability of response being considerably more impressive than previously observed with other agents not producing MRD-negative CRs. This perspective provides a view of where these agents may take us in the future as CLL therapy evolves with this exciting new class of drugs.


Melatonin prevents neuroinflammation and relieves depression by attenuating autophagy impairment through FOXO3a regulation.
Major depressive disorder (MDD) is a life-threatening illness characterized by mood changes and high rates of suicide. Although the role of neuroinflammation in MMD has been studied, the mechanistic interplay between antidepressants, neuroinflammation, and autophagy is yet to be investigated. The present study investigated the effect of melatonin on LPS-induced neuroinflammation, depression, and autophagy impairment. Our results showed that in mice, lipopolysaccharide (LPS) treatment induced depressive-like behaviors and caused autophagy impairment by dysregulating ATG genes. Moreover, LPS treatment significantly increased the levels of cytokines (TNFα, IL-1β, IL-6), enhanced NF-ᴋB phosphorylation, caused glial (astrocytes and microglia) cell activation, dysregulated FOXO3a expression, increased the levels of redox signaling molecules such as ROS/TBARs, and altered expression of Nrf2, SOD2, and HO-1. Melatonin treatment significantly abolished the effects of LPS, as demonstrated by improved depressive-like behaviors, normalized autophagy-related gene expression, and reduced levels of cytokines. Further, we investigated the role of autophagy in LPS-induced depressive-like behavior and neuroinflammation using autophagy inhibitors 3-MA and Ly294002. Interestingly, inhibitor treatment significantly abolished and reversed the anti-depressive, pro-autophagy, and anti-inflammatory effects of melatonin. The present study concludes that the anti-depressive effects of melatonin in LPS-induced depression might be mediated via autophagy modulation through FOXO3a signaling.


[Terminal phalangeal osteosclerosis in rheumatoid arthritis].
Osteosclerosis of the terminal phalanges of the hand has been reported in the collagen diseases including rheumatoid arthritis (RA). The current study was made to clarify the significance of terminal phalangeal osteosclerosis in RA. Hand x-ray films of 108 patients (male 13, female 95) with RA for more than 10 years were assessed by Halim's grading. Grade 2, (Distance between both cortices is < 1 mm but there is still an intermediate space discernible), and grade 3, (Complete fusion of both cortices, but the base of the terminal phalanx is open), were classified as the positive group. Grades 0 and 1 were classified as the negative group. Both groups were compared and sigma GS/D values was measured at terminal phalanges, middle phalanges and metacarpuses of right middle fingers using digital image processing method. sigma GS/D index was calculated as being equal to the ratio of sigma GS/D value of the terminal or middle phalanx to sigma GS/D value of the metacarpus. Thirty cases (28%) were positive (male 1, female 29). Osteosclerosis of terminal phalanges usually appeared in more than two digits bilaterally. It was located as follows; thumb-1 case, index-13 cases, middle-24 cases, ring-51 cases, little-58 cases. Obliteration of the medullary space of middle phalanges was seen in 5 cases. No difference was observed between the positive and negative groups about sex, age, the mean value of C-reactive protein, the incidence of Steinblocker's class classification and the rate of seropositive patients. Osteosclerosis of terminal phalanges was observed at first consultation in 93% of positive cases. Osteosclerosis is expected to appear in the early phase of RA and to last for more than 10 years. sigma GS/D index of the positive group (1.03 +/- 0.24, mean +/- SD) was greater than that of the negative group (0.63 +/- 0.25) (p < 0.05).


Why are you eating, mom? Maternal emotional, restrained, and external eating explaining children's eating styles.
This study investigated if three maternal eating styles (emotional eating style, external eating style, and restrained eating style) predict respective eating styles in children. In particular, we tested if these associations are different in mother-daughter dyads, compared to mother-son dyads. Data were collected twice, at the baseline (Time 1; T1) and at the 10-month follow-up (Time 2; T2), with N = 822 mother-child dyads participating at T1. Children (55% girls, 5-12 years old, M = 8.21, SD = 1.40) were interviewed; mothers (aged 23-59 years old, M = 35.93, SD = 5.24) completed the questionnaire assessing their eating styles. Participants' weight and height were measured objectively. Path analysis, accounting for dyadic interdependency and autocorrelations, was applied. In mother-daughter dyads, maternal emotional eating (T1) predicted daughters' emotional eating (T2) whereas maternal restrained eating (T1) predicted daughters' restrained eating (T2). There were no effects of external eating in mother-daughter dyads. A different pattern of associations was found for mother-son dyads, with maternal emotional eating (T1) and external eating (T1) predicting sons' emotional eating (T2) and external eating (T2), respectively. There was no effect of maternal restrained eating in mother-son dyads. Maternal eating styles explain child's eating styles with distinct effects depending on child's sex. Educating mothers about the effects of their own eating styles on daughters' and sons' eating styles might be useful to promote adequate responses to hunger and satiety signals.


Motor learning without physical practice: The effects of combined action observation and motor imagery practice on cup-stacking speed.
In this study we explored training effects for combined action observation and motor imagery (AO + MI) instructions on a complex cup-stacking task, without physical practice. Using a Graeco-Latin Square design, we randomly assigned twenty-six participants into four groups. This counterbalanced the within-participant factor of practice condition (AO + MI, AO, MI, Control) across four cup-stacking tasks, which varied in their complexity. On each of the three consecutive practice days participants experienced twenty trials under each of the three mental practice conditions. On each trial, a first-person perspective video depicted bilateral cup-stacking performed by an experienced model. During AO, participants passively observed this action, responding only to occasional colour cues. For AO + MI, participants imagined performing the observed action and synchronised their concurrent MI with the display. For MI, a sequence of pictures cued imagery of each stage of the task. Analyses revealed a significant main effect of practice condition both at the 'surprise' post-test (Day 3) and at the one-week retention test. At both time points movement execution times were significantly shorter for AO + MI compared with AO, MI and the Control. Execution times were also shorter overall at the retention compared with the post-test. These results demonstrate that a complex novel motor task can be acquired without physical training. Practitioners can therefore use AO + MI practice to supplement physical practice and optimise skill learning.


Allele-specific quantification of human leukocyte antigen transcript isoforms by nanopore sequencing.
While tens of thousands of HLA alleles have been identified by DNA sequencing, the contribution of alternative splicing to HLA diversity is not well characterized. In this study, we sought to determine if long-read sequencing could be used to accurately quantify allele-specific HLA transcripts in primary human lymphocytes. cDNA libraries were prepared from peripheral blood lymphocytes from 12 donors and sequenced by nanopore long-read sequencing. HLA reads were aligned to donor-specific reference sequences based on the known type of each donor. Allele-specific exon utilization was calculated as the proportion of reads aligning to each allele containing known exons, and transcript isoforms were quantified based on patterns of exon utilization within individual reads. Splice variants were rare among class I HLA genes (median exon retention rate 99%-100%), except for several HLA-C alleles with exon 5 spliced out of up to 15% of reads. Splice variants were also rare among class II HLA genes (median exon retention rate 98%-100%), except for HLA-DQB1. Consistent with previous work, exon 5 of HLA-DQB1 was spliced out in alleles with a mutated splice acceptor site at rs28688207. Surprisingly, a 28% loss of exon 5 was also observed in HLA-DQB1 alleles with an intact splice acceptor site at rs28688207. We describe a simple bioinformatic workflow to quantify allele-specific expression of HLA transcript isoforms. Further studies are warranted to characterize the repertoire of HLA transcripts expressed in different cell types and tissues across diverse populations.


Carbon Nanodot-Sensitized Modulation of Alzheimer's β-Amyloid Self-Assembly, Disassembly, and Toxicity.
The self-assembly of amyloidogenic peptides into β-sheet-rich aggregates is a general feature of many neurodegenerative diseases, including Alzheimer's disease, which signifies the need for the effective attenuation of amyloid aggregation toward alleviating amyloid-associated neurotoxicity. This study reports that photoluminescent carbon nanodots (CDs) can effectively suppress Alzheimer's β-amyloid (Aβ) self-assembly and function as a β-sheet breaker disintegrating preformed Aβ aggregates. This study synthesizes CDs using ammonium citrate through one-pot hydrothermal treatment and passivates their surface with branched polyethylenimine (bPEI). The bPEI-coated CDs (bPEI@CDs) exhibit hydrophilic and cationic surface characteristics, which interact with the negatively charged residues of Aβ peptides, suppressing the aggregation of Aβ peptides. Under light illumination, bPEI@CDs display a more pronounced effect on Aβ aggregation and on the dissociation of β-sheet-rich assemblies through the generation of reactive oxygen species from photoactivated bPEI@CDs. The light-triggered attenuation effect of Aβ aggregation using a series of experiments, including photochemical and microscopic analysis, is verified. Furthermore, the cell viability test confirms the ability of photoactivated bPEI@CDs for the suppression of Aβ-mediated cytotoxicity, indicating bPEI@CDs' potency as an effective anti-Aβ neurotoxin agent.


Regional and age-related differences in mRNA composition of intracellular Ca(2+)-release channels of rat cardiac myocytes.
We investigated the mRNA distribution of three different ryanodine receptors (RyR) and of the intracellular Ca(2+)-release channel/inositol 1,4,5-trisphosphate receptor (IP3R) type 1 in the rat heart during development and aging. In situ hybridization analysis shows that RyR1 mRNA is never expressed in the heart at any of the stages examined: RyR2 mRNA is detectable in cardiomyocytes in the early embryonic stages, whereas RyR3 mRNA accumulates in cardiomyocytes around birth. IP3R mRNA appears at first in the primitive atrium at embryonic day 11 and in subsequent stages it is detectable also in a minor population of ventricular myocytes, which presumably correspond to conduction system precursors. In the adult heart, no apparent difference in hybridization signal intensity is observed between atrial and ventricular working myocytes either with RyR2, RyR3 or IP3R cRNA probes, except for myocytes of the heart conduction system, which differ from working myocytes in the intensity of the hybridization signals for each probe. Additional differences are detected in the senescent heart with the IP3R cRNA probe, which hybridizes with atrial myocytes stronger than with ventricular ones. RNase protection analysis confirms the temporal differences in RyR2 and RyR3 transcript accumulation observed during heart development and reveals a significant increase of IP3R mRNA in the atrial myocardium during aging. Thus, the composition of intracellular Ca(2+)-release channel mRNAs of the rat heart shows temporal and regional variations: such changes might reflect important differences in transcriptional regulation of these genes among myocytes.


Atom-Pair Kinetics with Strong Electric-Dipole Interactions.
Rydberg-atom ensembles are switched from a weakly to a strongly interacting regime via adiabatic transformation of the atoms from an approximately nonpolar into a highly dipolar quantum state. The resultant electric dipole-dipole forces are probed using a device akin to a field ion microscope. Ion imaging and pair-correlation analysis reveal the kinetics of the interacting atoms. Dumbbell-shaped pair-correlation images demonstrate the anisotropy of the binary dipolar force. The dipolar C_{3} coefficient, derived from the time dependence of the images, agrees with the value calculated from the permanent electric-dipole moment of the atoms. The results indicate many-body dynamics akin to disorder-induced heating in strongly coupled particle systems.


Effect of gravity change on the production of thrombopoietic growth factors.
It is reported that the stay in the space develops anemia, thrombocytopenia, and altered function and structure of red blood cell. The mechanism of these abnormalities was not clarified yet. The cloning of the thrombopoietin (TPO), followed by the analysis of TPO and c-mpl (its cellular receptor) knockout mice confirmed its role as the primary regulator of thrombopoiesis. TPO has been shown to stimulate both megakaryocyte colony growth from marrow progenitor cells and the maturation of immature megakaryocyte to form functional platelet. This process includes the massive cytoskeletal rearrangement, such as proplatelet formation and fragmentation of proplatelet. In this study we have focused on the production of thrombopoietic growth factors in mice those were exposed to gravity change by parabolic flight (PF).


Identification of gene expression logical invariants in Arabidopsis.
Numerous gene expression datasets from diverse tissue samples from the plant variety Arabidopsis thaliana have been already deposited in the public domain. There have been several attempts to do large scale meta-analyses of all of these datasets. Most of these analyses summarize pairwise gene expression relationships using correlation, or identify differentially expressed genes in two conditions. We propose here a new large scale meta-analysis of the publicly available Arabidopsis datasets to identify Boolean logical relationships between genes. Boolean logic is a branch of mathematics that deals with two possible values. In the context of gene expression datasets we use qualitative high and low expression values. A strong logical relationship between genes emerges if at least one of the quadrants is sparsely populated. We pointed out serious issues in the data normalization steps widely accepted and published recently in this context. We put together a web resource where gene expression relationships can be explored online which helps visualize the logical relationships between genes. We believe that this website will be useful in identifying important genes in different biological context. The web link is http://hegemon.ucsd.edu/plant/.


Comparing atmospheric and hypoxic cultured mesenchymal stem cell transcriptome: implication for stem cell therapies targeting intervertebral discs.
Mesenchymal stem cells (MSCs) represent an attractive avenue for cellular therapies targeting degenerative diseases. MSC in vitro expansion is required in order to obtain therapeutic numbers during the manufacturing process. It is known that culture conditions impact cellular properties and behavior after in vivo transplantation. In this study, we aimed at evaluating the benefit of hypoxic culturing of human bone marrow derived mesenchymal stem cells on cell fitness and whole genome expression and discussed its implication on cellular therapies targeting orthopedic diseases such as chronic lower back pain. Human bone marrow mesenchymal stem cells (hBMMSCs) were isolated from fresh human anticoagulated whole bone marrow and were cultured side by side in atmospheric (20% O2) and hypoxic (5% O2) oxygen partial pressure for up to 3 passages. Stem cell fitness was assessed by clonogenic assay, cell surface marker expression and differentiation potential. Whole genome expression was performed by mRNA sequencing. Data from clonogenic assays, cell surface marker by flow cytometry and gene expression by quantitative PCR were analyzed by two-tailed paired Student's t-test. Data from mRNA sequencing were aligned to hg19 using Tophat-2.0.13 and analyzed using Cufflinks-2.1.1. Hypoxic culturing of hBMMSCs had positive effects on cell fitness, as evidenced by an increased clonogenicity and improved differentiation potential towards adipocyte and chondrocyte lineages. No difference in osteoblast differentiation or in cell surface markers were observed. Only a small subset of genes (34) were identified by mRNA sequencing to be significantly dysregulated by hypoxia. When clustered by biological function, these genes were associated with chondrogenesis and cartilage metabolism, inflammation and immunomodulation, cellular survival, migration and proliferation, vasculogenesis and angiogenesis. Hypoxic culturing positively impacted hBMMSCs fitness and transcriptome, potentially improving inherent properties of these cells that are critical for the development of successful cellular therapies. Hypoxic culturing should be considered for the in vitro expansion of hBMMSCs during manufacturing of cellular therapies targeting orthopedic disorders such as lower back pain.


Field rickets in turkeys: relationship to vitamin D.
Thirty-two outbreaks of leg disorders in turkeys were investigated during 1981-1985. Among them, 22 were characterized by a low percentage of bone ash and were considered as field rickets. Most of the field rickets cases exhibited reduced plasma calcium and inorganic phosphorus. Plasma 25-hydroxyvitamin D3 and intestinal calcium-binding protein were lower in the rachitic than in normal turkeys. These symptoms are typical of rickets resulting from vitamin D deficiency. The vitamin D3 equivalence of a diet that had been fed during field rickets outbreaks was assayed biologically and found to be 111 micrograms/kg diet, about eight times the minimal requirement. In two other cases no symptoms of rickets were observed in turkeys fed diets that had been previously consumed during field rickets outbreaks. The results indicate that in some cases of field rickets there was no involvement of dietary factors and confirm a previous suggestion that field rickets may result from defects in metabolism of vitamin D3, or in its expression.


FOXD1-AS1 upregulates FOXD1 to promote oral squamous cell carcinoma progression.
Recently, increasing attention has been concentrated on decrypting the potential of long non-coding RNAs (lncRNAs) in influencing the progression of human tumors, oral squamous cell carcinoma (OSCC) included. The role of a novel lncRNA, forkhead box D1 antisense RNA 1 (FOXD1-AS1), has been discussed in multiple cancers. Nevertheless, its function and relevant mechanism in OSCC have been not probed yet. FOXD1-AS1 expression was detected via RT-qPCR. Colony formation, EdU, transwell and Western blot analyses tested the functional role of FOXD1-AS1 in OSCC cells. The relationship between RNAs was assessed by a series of mechanical assays. FOXD1-AS1 was expressed at a high level in head and neck squamous cell carcinoma (HNSC). Knockdown of FOXD1-AS1 exerted repressive impacts on OSCC cell proliferation, migration, invasion, and EMT. Moreover, FOXD1-AS1 positively regulated its nearby gene FOXD1 via interacting with miR-369-3p. In addition, adenosine deaminase RNA specific (ADAR), known as a RNA-binding protein (RBP), was capable to bind with FOXD1-AS1 and FOXD1 simultaneously, and could regulate the stability of FOXD1 mRNA. Aside from that, rescue assays delineated that FOXD1-AS1 promoted OSCC progression via upregulating FOXD1. FOXD1-AS1 elevates FOXD1 expression to promote OSCC malignant phenotypes through miR-369-3p and ADAR.


Effect of endodontic irrigation with 1% sodium hypochlorite and 17% EDTA on primary teeth: a scanning electron microscope analysis.
This study evaluated the efficacy of 2 final irrigation solutions for removal of the smear layer (SL) from root canals of primary teeth, using scanning electron microscope (SEM) analysis. Thirty primary molars were selected and a single operator instrumented the canals. The initial irrigation was done with a 1% sodium hypochlorite (NaOCl) solution. After the preparation, the roots were randomly divided into 3 groups for final irrigation: Group 1, 1% NaOCl (n = 10); Group 2, 17% EDTA + 1% NaOCl (n = 10); and Group 3, 17% EDTA + saline solution (n = 10). The roots were prepared for SEM analysis (magnification 1000X). The photomicrographs were independently analyzed by 2 investigators with SEM experience, attributing scores to each root third in terms of SL removal. Kruskal-Wallis and Mann-Whitney tests revealed that there was no statistical difference between the groups (P = 0.489). However, a statistical difference was found (P < 0.05) in a comparison of root thirds, with the apical third having the worst results. Comparing the thirds within the same group, all canals showed statistical differences between the cervical and apical thirds (P < 0.05). The authors determined that no substance or association of substances were able to completely remove SL.


Postural hypoxemia in the postpneumonectomy patient.
A patient with platypnea and orthodeoxia after right pneumonectomy is described. Cardiac catheterization revealed arterial desaturation in the upright position associated with increased right to left intracardiac shunting through a patent foraman ovale. The absence of a pressure gradient between the left and right atrium in either the supine or sitting position indicated that the orthodeoxia resulted from an increase in streaming of blood from the inferior vena cava to the left atrium caused by mechanical distortion of the interatrial septum.


[Telomerase activity of tumors in the head and neck].
Telomerase is a maintaining enzyme of telomere repeats. Telemorase is expressed in immortal and cancer cells, but not in the normal tissues. We investigated telomerase activity of tumors in the head and neck. Telomerase activity was found in all of the cell lines derived from squamous cell carcinomas of the head and neck. Six of 9 (67%) primary fresh tissues from squamous cell carcinomas of the head and neck contained detectable telomerase activity. There were no apparent relationship between telomerase activity and clinical stage. Telomerase activity differed, depend on the responses to chemotherapy and radiotherapy: complete response, 0 of 3(0%); partial response; 0 of 1(0%); no change; 7 of 7 (100%). Therapy-resistant tumor seemed to have much telomerase activity and much malignant potential. In benign tumors, neither of 2 benign parotid tumors expressed telomerase activity, but one of 2(50%)papillomas had the telomerase activity. Telomerase is considered useful tumor marker for the diagnosis and prognosis.


The analytical process to search for metabolomics biomarkers.
The scant number of available metabolomics biomarkers does not reflect the extent of the research in this field. Looking for the reasons of failure, the authors, as analytical chemists, critically discuss each of the steps in the analytical process that requires improvements. They find insufficient information about how the experimental part is developed. After revising the steps from sampling to obtainment of the analytical sample (from typical samples such as blood and urine to others less common such as sweat or saliva), the need for data and metadata for either reproduction of a given study or for taking the study as starting point after biomarker discovery is criticized. The separation and analysis steps are also revised as does data treatment. After the sources of errors from the analytical process are overcome, subsequent steps in the implementation of biomarkers to reach the final aim of clinical adoption should be supported as required.


Switching and growth for microbial populations in catastrophic responsive environments.
Phase variation, or stochastic switching between alternative states of gene expression, is common among microbes, and may be important in coping with changing environments. We use a theoretical model to assess whether such switching is a good strategy for growth in environments with occasional catastrophic events. We find that switching can be advantageous, but only when the environment is responsive to the microbial population. In our model, microbes switch randomly between two phenotypic states, with different growth rates. The environment undergoes sudden catastrophes, the probability of which depends on the composition of the population. We derive a simple analytical result for the population growth rate. For a responsive environment, two alternative strategies emerge. In the no-switching strategy, the population maximizes its instantaneous growth rate, regardless of catastrophes. In the switching strategy, the microbial switching rate is tuned to minimize the environmental response. Which of these strategies is most favorable depends on the parameters of the model. Previous studies have shown that microbial switching can be favorable when the environment changes in an unresponsive fashion between several states. Here, we demonstrate an alternative role for phase variation in allowing microbes to maximize their growth in catastrophic responsive environments.


Development of PBC/SSc overlap syndrome in chronic GVHD patient: immunological implications in the presence of mitochondrial, nucleolar and spindle midzone autoantigens.
Chronic Graft versus Host Disease (cGVHD) is a complex disease resulting from donor T-cell recognition of a genetically disparate recipient that is unable to reject donor cells after allogeneic Stem Cell Transplantation (HSCT). cGVHD has some features resembling to autoimmune diseases (AD) such as Sjögren syndrome, primary biliary cirrhosis (PBC) and scleroderma (SSc). Also patients with cGVHD could develop extensive cGVHD with scleroderma-like skin manifestations and other clinical signs similar to those of patients with scleroderma. We take into consideration a patient with GVHD that developed PBC/SSc overlap syndrome with a complex and particular autoantibodies profile. Indirect immunofluorescence (IIF) with double coloration showed a cytoplasmic mitochondrial-like pattern, a clumpy nucleolar staining pattern, and a cell-cycle related staining pattern. Following anaphase onset, proteins regulator of cytokinesis localizes to the overlap zone on the ends of midzone microtubules and becomes compacted during furrow ingression to form the midbody. Second level tests confirmed the presence of anti-mitochondrial antibodies M2-subunit but no other autoantibodies were found. We performed a home-made immunoblot analysis that identified a 37 kDa fibrillarin band, and not identify 47 kDa, 31KDa and 18/20 kDa bands. After literature review of these possible cellular localizations, the proteins recognized by our patient's serum seem likely to be Aab to core midzone organizer components. However, due to the unavailability of the proper techniques in our laboratory, we were not able to further characterize them. The pathogenesis and morbidity of cGVHD after HSCT remains enigmatic, but the presence of specific autoantibodies are the hallmark of AD and represent a possibility of differential diagnosis. Standard techniques combined with the use of non-routinely laboratory techniques are a usefully and complementary method for studying difficult and particular cases. In fact, these autoantibodies will be considered as ''diagnostic'' and not as ''esoteric'' antibodies. In conclusion, a re-assessment of the diagnostic protocols in cGVHD together with a precise observation of the clinical and laboratory picture will ultimately help us clarify the disease and could provide a better understanding of the immune network deregulation.


Magnetic resonance imaging of distal sesamoidean ligament injury.
Distal sesamoidean ligament injury is a recognized cause of lameness but diagnosis using ultrasonography is sometimes difficult. Herein, we describe the normal appearance of the distal sesamoidean ligaments on magnetic resonance (MR) images and the changes that occur when the ligaments are injured. The appearance of the distal sesamoidean ligaments on MR images from 66 control horses and 58 horses with distal sesamoidean desmitis were described and the cross-sectional area and signal intensity of the ligaments measured. In control horses, the ligaments had a characteristic appearance and strong left-right symmetry, and the lateral oblique sesamoidean ligament was larger and had higher signal intensity than the medial ligament. Cross-sectional area and signal intensity were significantly greater in injured straight sesamoidean ligaments compared with the controls. Signal intensity increased significantly with oblique sesamoidean desmitis compared with the controls. Lesions of the distal sesamoidean ligaments were considered the sole cause of lameness in only 2 of 58 horses. Eighty percent of lesions in the distal sesamoidean ligaments were not detected using ultrasonography.


Synergistic Cytotoxicity of Lenalidomide and Dexamethasone in Mantle Cell Lymphoma via Cereblon-dependent Targeting of the IL-6/STAT3/PI3K Axis.
At our center, relapsed mantle cell lymphoma (MCL) can be treated with maintenance therapy composed of consecutive low-dose lenalidomide and short-term, high-dose dexamethasone (LD regimen), which achieves good responses (longer overall survival and progression-free survival) and low toxicity. Cereblon is probably targeted by both lenalidomide and dexamethasone, which leads to synergistic cytotoxicity in MCL by inhibiting the interleukin-6/signal transducer and activator of transcription 3 (IL-6/STAT3), phosphatidylinositol 3-kinase (PI3K)/AKT and AKT2/Forkhead box O3 (FOXO3A)/BCL2-like 11 (BIM) pathways. The two drugs synergistically inhibit the same pathways, but through different sites. Cereblon was found expressed in most of the MCL tissues (91.3% positivity). Moreover, cereblon expression is positively correlated with LD regimen sensitivity: long-term lenalidomide exposure downregulates cereblon and induces multi-drug resistance against lenalidomide, dexamethasone, cytarabine, cisplatin, and methotrexate in vitro. Removal of lenalidomide resensitizes lenalidomide-resistant MCL cells to lenalidomide and dexamethasone. Our work suggests that rotating the LD regimen with other regimens would improve MCL maintenance therapy.


Evaluation of K-ras and p53 expression in pancreatic adenocarcinoma using the cancer genome atlas.
Genetic alterations in K-ras and p53 are thought to be critical in pancreatic cancer development and progression. However, K-ras and p53 expression in pancreatic adenocarcinoma have not been systematically examined in The Cancer Genome Atlas (TCGA) Data Portal. Information regarding K-ras and p53 alterations, mRNA expression data, and protein/protein phosphorylation abundance was retrieved from The Cancer Genome Atlas (TCGA) databases, and analyses were performed by the cBioPortal for Cancer Genomics. The mutual exclusivity analysis showed that events in K-ras and p53 were likely to co-occur in pancreatic adenocarcinoma (Log odds ratio = 1.599, P = 0.006). The graphical summary of the mutations showed that there were hotspots for protein activation. In the network analysis, no solid association between K-ras and p53 was observed in pancreatic adenocarcinoma. In the survival analysis, neither K-ras nor p53 were associated with both survival events. As in the data mining study in the TCGA databases, our study provides a new perspective to understand the genetic features of K-ras and p53 in pancreatic adenocarcinoma.


Biodiversity vs. biocontrol: positive and negative effects of alternative prey on control of slugs by carabid beetles.
Environment-friendly farming techniques seek to increase invertebrate biodiversity in part with the intention of encouraging greater numbers of predators that will help to control crop pests. However, in theory, this effect may be negated if the availability of a greater abundance and diversity of alternative prey diverts predators away from feeding on pests. The hypothesis that access to alternative prey can lead to reduced pest suppression under semi-field conditions was tested. Alternative prey type and diversity were manipulated in 70 mesocosms over 7+ weeks in the presence of the carabid Pterostichus melanarius (Illiger), a known predator of slugs, and reproducing populations of the slug Deroceras reticulatum (Müller). Significantly fewer slugs survived where no alternative prey were provided. Maximum slug numbers and biomass were found in treatments containing either carabids plus a high diversity of alternative prey (many species of earthworm and three of Diptera larvae) or a single additional prey (blowfly larvae, Calliphora vomitoria Linnaeus). In these treatments slug numbers and biomass were as high as in plots lacking predators. The effects of alternative prey were taxon-specific. Alternative prey strongly affected carabid fitness in terms of biomass and egg load. The fittest predators (those with access to high alternative prey diversity or C. vomitoria larvae) reduced slug numbers the least. The mean individual slug weights were greater in treatments with alternative prey than where no alternative prey were provided to the carabids. These results suggest that pests may survive and reproduce more rapidly in patches where predators have access to alternative prey.


Extracellular ATP plays an important role in systemic wound response activation.
Mechanical wounding occurs in plants during biotic or abiotic stresses and is associated with the activation of long-distance signaling pathways that trigger wound responses in systemic tissues. Among the different systemic signals activated by wounding are electric signals, calcium, hydraulic, and reactive oxygen species (ROS) waves. The release of glutamate (Glu) from cells at the wounded tissues was recently proposed to trigger systemic signal transduction pathways via GLU-LIKE RECEPTORs (GLRs). However, the role of another important compound released from cells during wounding (extracellular ATP [eATP]) in triggering systemic responses is not clear. Here, we show in Arabidopsis (Arabidopsis thaliana) that wounding results in the accumulation of nanomolar levels of eATP and that these levels are sufficient to trigger the systemic ROS wave. We further show that the triggering of the ROS wave by eATP during wounding requires the PURINORECEPTOR 2 KINASE (P2K) receptor. Application of eATP to unwounded leaves triggered the ROS wave, and the activation of the ROS wave by wounding or eATP application was suppressed in mutants deficient in P2Ks (e.g. p2k1-3, p2k2, and p2k1-3p2k2). In addition, expression of systemic wound response (SWR) transcripts was suppressed in mutants deficient in P2Ks during wounding. Interestingly, the effect of Glu and eATP application on ROS wave activation was not additive, suggesting that these two compounds function in the same pathway to trigger the ROS wave. Our findings reveal that in addition to sensing Glu via GLRs, eATP sensed by P2Ks plays a key role in the triggering of SWRs in plants.


gdnf affects early diencephalic dopaminergic neuron development through regulation of differentiation-associated transcription factors in zebrafish.
Glial cell line-derived neurotrophic factor (GDNF) has been reported to enhance dopaminergic neuron survival and differentiation in vitro and in vivo, although those results are still being debated. Glial cell line-derived neurotrophic factor (gdnf) is highly conserved in zebrafish and plays a role in enteric nervous system function. However, little is known about gdnf function in the teleost brain. Here, we employed clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 to impede gdnf function in the maintenance of dopaminergic neuron development. Genotyping of gdnf crispants revealed successful deletions of the coding region with various mutant band sizes and down-regulation of gdnf transcripts at 1, 3 and 7 day(s) post fertilization. Notably, ~20% reduction in ventral diencephalic dopaminergic neuron numbers in clusters 8 and 13 was observed in the gdnf-deficient crispants. In addition, gdnf depletion caused a modest reduction in dopaminergic neurogenesis as determined by 5-ethynyl-2'-deoxyuridine pulse chase assay. These deleterious effects could be partly attributed to deregulation of dopaminergic neuron fate specification-related transcription factors (otp,lmx1b,shha,and ngn1) in both crispants and established homozygous mutants with whole mount in-situ hybridization (WISH) on gdnf mutants showing reduced otpb and lmx1b.1 expression in the ventral diencephalon. Interestingly, locomotor function of crispants was only impacted at 7 dpf, but not earlier. Lastly, as expected, gdnf deficiency heightened crispants vulnerability to 1-methyl-4-phenylpyridinium toxic insult. Our results suggest conservation of teleost gdnf brain function with mammals and revealed the interactions between gdnf and transcription factors in dopaminergic neuron differentiation.


Beliefs and attitudes about addressing alcohol consumption in health care: a population survey in England.
Despite robust evidence for their effectiveness, it has proven difficult to translate alcohol prevention activities into routine health care practice. Previous research has identified numerous provider-level barriers affecting implementation, but these have been less extensively investigated in the wider population. We sought to: (1) investigate patients' beliefs and attitudes to being asked about alcohol consumption in health care; and (2) identify the characteristics of those who are supportive of addressing alcohol consumption in health care. Cross-sectional household interviews conducted as part of the national Alcohol Toolkit Study in England between March and April 2017. Data were collected on age, gender, social grade, drinking category, and beliefs and attitudes to being asked about alcohol in routine health care. Unadjusted and multivariate-adjusted logistic regression models were performed to investigate associations between socio-demographic characteristics and drinking category with being "pro-routine" (i.e. 'agree completely' that alcohol consumption should be routinely addressed in health care) or "pro-personal" (i.e. 'agree completely' that alcohol is a personal matter and not something health care providers should ask about). Data were collected on 3499 participants, of whom 50% were "pro-routine" and 10% were "pro-personal". Those in social grade C1, C2, D and E were significantly less likely than those in AB of being "pro-routine". Women were less likely than men to be "pro-personal", and those aged 35-44 or 65 years plus more likely to be "pro-personal" compared with participants aged 16-24. Respondents aged 65 plus were twice as likely as those aged 16-24 to agree completely that alcohol consumption is a personal matter and not something health care providers should ask about (OR 2.00, 95% CI 1.34-2.99). Most adults in England agree that health care providers should routinely ask about patients' alcohol consumption. However, older adults and those in lower socio-economic groups are less supportive. Drinking status appears to have limited impact on whether people believe that alcohol is a personal matter and not something health care providers should ask about. Open Science Framework ( https://osf.io/xn2st/ ).


Three-body recombination at large scattering lengths in an ultracold atomic gas.
We study three-body recombination in an optically trapped ultracold gas of cesium atoms with precise magnetic control of the s-wave scattering length a. At large positive values of a, we measure the dependence of the rate coefficient on a and confirm the theoretically predicted scaling proportional to a(4). Evidence of recombination heating indicates the formation of very weakly bound molecules in the last bound energy level.


Creating safer health systems: Lessons from other sectors and an account of an application in the Safer Clinical Systems programme.
Despite well meaning initiatives over decades, the percentage of inpatients suffering adverse events remains constant in most advanced health systems. The notion of incident reporting as used in other safety critical industries has proved far less effective in healthcare. This article describes a new patient safety paradigm in the search for improved patient safety in healthcare. Underpinned by a holistic use of human factors the Safer Clinical Systems programme involves a proactive, risk-based approach seeking to eliminate or control risk before it is converted to patient harm. The tools and techniques applied by healthcare professional in real-life settings are described along with the outcomes of a significant reduction in risk and improvement in safety culture as measured by the Safety Culture Index. The challenges of applying the approach are discussed but it is argued that important progress could be made if a critical mass of healthcare staff were helped to acquire skills in human factors.


Leukocyte transmigration into tissue-engineered constructs is influenced by endothelial cells through Toll-like receptor signaling.
Inflammation plays a crucial role in tissue regeneration, wound healing, and the success of tissue-engineered constructs. The aim of this study was to investigate the influence of human umbilical vein endothelial cells (ECs) on leukocyte transmigration when co-cultured with primary human bone marrow-derived multipotent stromal cells (MSCs). MSCs with and without ECs were cultured in poly (L-lactide-co-1, 5-dioxepan-2-one) (poly (LLA-co-DXO)) scaffolds for 1 week in vitro in a bioreactor system, after which they were implanted subcutaneously in non-obese diabetic/severe combined immunodeficient mice. After 1 and 3 weeks, scaffolds were retrieved, and the mRNA expression of interleukin 1-beta (IL-1β), IL-6, IL-10, hypoxia-inducible factor 1-alpha (HIF-1α), HIF-1β, and mammalian target of rapamycin was examined by real-time reverse transcription-polymerase chain reaction. Furthermore, immunofluorescent staining was performed for IL-1β, IL-6, neutrophils, and CD11b. In addition, Western blotting was done for IL-1β and IL-6. Leukocyte transmigration genes and genes in Toll-like receptor pathways, expressed by MSCs cultured in vitro with or without ECs, were further investigated with a microarray dataset. In vitro, genes involved in leukocyte transmigration and Toll-like receptor pathways were clearly influenced by the addition of ECs. Platelet/endothelial cell adhesion molecule-1 (PECAM-1) and cadherin-5 (CDH5), both genes involved in leukocyte transmigration, were expressed significantly higher in the MSC/EC group. The recruitment of leukocytes into tissue-engineered constructs with MSCs is strongly influenced by the addition of ECs via activation of leukocyte transmigration and Toll-like receptor pathways.


S-allele diversity in Sorbus aucuparia and Crataegus monogyna (Rosaceae: Maloideae).
RT-PCR was used to obtain the first estimates from natural populations of allelic diversity at the RNase-based gametophytic self-incompatibility locus in the Rosaceae. A total of 20 alleles were retrieved from 20 Sorbus aucuparia individuals, whereas 17 alleles were found in 13 Crataegus monogyna samples. Estimates of population-level allele numbers fall within the range observed in the Solanaceae, the only other family with RNase-based incompatibility for which estimates are available. The nucleotide diversity of S-allele sequences was found to be much lower in the two Rosaceae species as compared with the Solanaceae. This was not due to a lower sequence divergence among most closely related alleles. Rather, it is the depth of the entire genealogy that differs markedly in the two families, with Rosaceae S-alleles exhibiting more recent apparent coalescence. We also investigated patterns of selection at the molecular level by comparing nucleotide diversity at synonymous and nonsynonymous sites. Stabilizing selection was inferred for the 5' region of the molecule, while evidence of diversifying selection was present elsewhere.


Experimental infection and horizontal transmission of Modoc virus in deer mice (Peromyscus maniculatus).
Deer mice (Peromyscus maniculatus) were inoculated with a sublethal dose of a field strain of Modoc virus to determine patterns of viral persistence, shedding, and transmission. Blood, serum, urine, fecal, and oral swab samples were collected at selected intervals until 63 days postinoculation (PI) after which lung, liver, spleen, kidney, and salivary glands were explanted. Viral assays were conducted by intracranial inoculations of suckling mice and antibody titers were determined by the micro-complement-fixation test. Viremias lasted for up to 4 days PI. Antibody titers were present by day 8 PI, peaked at day 13-20 PI, and persisted until day 63 PI. There was no evidence of viral shedding in urine, fecal, or oral swab samples. Virus was detected in explanted lungs only. In a separate experiment, deer mice were inoculated with virus and lungs were removed from five mice per wk for 10 wk. Indirect fluorescent antibody (IFA) techniques were used to determine the location of virus in lung tissue and to examine fixed tissue for lesions. IFA showed virus in lung parenchymal cells beginning 42 days PI and persisting at least 70 days PI. No histopathologic changes were seen. Horizontal transmission of the virus was studied by placing uninoculated mice with inoculated mice for 42 days and determining if the test animals developed antibodies or had virus in their lungs. Fifty-percent of the uninoculated mice developed antibody. One of these animals had virus in its lungs. Therefore, Modoc virus may be transmitted by direct contact.


Laparoscopic-assisted bowel resection in pediatric/adolescent inflammatory bowel disease: laparoscopic bowel resection in children.
The purpose of this study is to discuss indications, technical approach, and morbidity of laparoscopic approaches to major bowel resection in the pediatric/adolescent population with inflammatory bowel disease and familial polyposis. Retrospective review of laparoscopic-assisted bowel procedures between May 1991 and January 2002 was performed. Laparoscopic-assisted bowel resection is defined as complete intracorporeal mobilization and devascularization of a segment of colon or rectum. The indications for extracorporeal vs. intracorporeal anastomosis will be discussed. Clinically unstable, septic, or massively bleeding patients were not candidates for this technique. The decision to attempt the laparoscopic approach was based on the experience of the consulting surgeon. There were 31 patients, including 14 females. Five patients had undergone prior surgery. Twenty-nine patients had inflammatory bowel disease, one had familial polyposis, and one had a cavernous hemangioma. We included all pediatric/adolescent patients in our practice undergoing laparoscopic resection. Twenty-nine patients had 33 laparoscopic operations, including proctocolectomy with ileal pouch-anal anastomosis (n = 14), proctocolectomy with ileostomy (n = 3), ileocolectomy with ileocolic anastomosis (n = 13), and small-bowel obstruction (n = 1). Average operating time was 158 (range, 30-400) minutes, with average blood loss of 159 ml. Average wound length was 4.9 cm. The complication rate was 16 percent (n = 5), with one anastomotic leak. The rate of conversion to open operations was 5.8 percent. Liquid diet was begun on Day 3, and the average length of stay was 5.9 days. Major laparoscopic bowel surgery can be performed safely in the pediatric/adolescent population, with reasonable operative times, low conversion to open operations, and low morbidity.


Third dose of BNT162b2 improves immune response in liver transplant recipients to ancestral strain but not Omicron BA.1 and XBB.
Vaccine immunogenicity in transplant recipients can be impacted by the immunosuppressive (IS) regimens they receive. While BNT162b2 vaccination has been shown to induce an immune response in liver transplant recipients (LTRs), it remains unclear how different IS regimens may affect vaccine immunogenicity after a third BNT162b2 dose in LTRs, which is especially important given the emergence of the Omicron sublineages of SARS-CoV-2. A total of 95 LTRs receiving single and multiple IS regimens were recruited and offered three doses of BNT162b2 during the study period. Blood samples were collected on days 0, 90, and 180 after the first BNT162b2 dose. At each time point, levels of anti-spike antibodies, their neutralizing activity, and specific memory B and T cell responses were assessed. LTRs receiving single IS regimens showed an absence of poor immunogenicity, while LTRs receiving multiple IS regimens showed lower levels of spike-specific antibodies and immunological memory compared to vaccinated healthy controls after two doses of BNT162b2. With a third dose of BNT162b2, spike-specific humoral, memory B, and T cell responses in LTR significantly improved against the ancestral strain of SARS-CoV-2 and were comparable to those seen in healthy controls who received only two doses of BNT162b2. However, LTRs receiving multiple IS regimens still showed poor antibody responses against Omicron sublineages BA.1 and XBB. A third dose of BNT162b2 may be beneficial in boosting antibody, memory B, and T cell responses in LTRs receiving multiple IS regimens, especially against the ancestral Wuhan strain of SARS-CoV-2. However, due to the continued vulnerability of LTRs to presently circulating Omicron variants, antiviral treatments such as medications need to be considered to prevent severe COVID-19 in these individuals.